1-2-dielaidoylphosphatidylethanolamine and Ovarian-Neoplasms

To study the inhibitory effect of endostatin mediated by lipofectin on transplanted ovarian cancer in nude mice.. Constructed recombinant vector pVAX1-sEn expressing human endostatin protein was transfected into ovarian cancer cell line 3AO by lipofectin. mRNA of endostatin was detected by RT-PCR. The expression of endostatin in supernatants was detected by enzyme-linked immunosorbent assay (ELISA). The inhibitory effect of pVAX1-sEn on endothelial cell line ECV-204 was detected by methyl thiazolyl tetrazolium (MTT). By use of lipofectin mediated pVAX1-sEn for intratumor injection, the inhibitory effect on growth of ovarian cancer was observed.. The result of RT-PCR showed there was a specific band at 610 bp. The expression quantity of endostatin in transfected cell supernatant was (201 +/- 8) ng/ml by ELISA. MTT showed pVAX1-sEn transfected cell supernatant could effectively inhibit the growth of ECV-204, the highest inhibitory ratio was 42%. The tumor volumes in pVAX1-sEn treatment group was (0.85 +/- 0.18) cm(3), significantly smaller than that in normal saline control group (1.90 +/- 0.28) cm(3) and pVAX1 control group (1.78 +/- 0.32) cm(3) (P < 0.05). HE stain in tumor tissue showed that there were obvious necrosis cells in the pVAX1-sEn treatment group, but there were flourishly growing tumor cells in pVAX1 and normal saline control groups.. pVAX1-sEn mediated by lipofectin can effectively inhibit the growth of ovarian cancer.

Topics: Animals; Cell Line, Tumor; Cell Proliferation; Down-Regulation; Endostatins; Female; Gene Expression; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Ovarian Neoplasms; Phosphatidylethanolamines; Random Allocation; Transfection

Viral vector systems are the most commonly used gene transfer tools for clinical gene therapy. However, lipofection systems are potential alternatives because of lower immunogenicity and easier cGMP production, but in vivo stability and transduction efficacy need to be improved. Therefore, we investigated gene transduction efficiency of our novel cGMP cationic lipids, CCQ22 and CCQ32, by FACS analysis. Toxicity analysis was performed to determine the cytotoxic side effects of the novel lipids. To evaluate the stability of the compounds in the context of local delivery to patients with intraperitoneally metastatic ovarian cancer, gene transfer was also tested in the presence of malignant ascites. Our novel cGMP standard lipids mediated gene transfer rates of more than 50%. However, for most cell lines cytotoxic side effects were similar to our reference lipofection system. In general, ascites had no major influence on gene transduction rates with the novel lipids. Our results suggest that CCQs may compare favorably with commercially available lipofection systems. These promising results facilitate further analysis of the compounds.

Topics: Ascites; Breast Neoplasms; Cell Line, Tumor; Cholesterol Esters; Female; Genetic Vectors; Humans; Lipids; Liposomes; Ovarian Neoplasms; Phosphatidylethanolamines; Plasmids; Transduction, Genetic

The cisplatin induced enhancement of in situ lipofection was optimized by considering the factors that can increase the degree of sensitization. Two other anticancer drugs, mechlorethamine (nitrogen mustard) and taxol, enhanced CAT gene expression but the degree of sensitization was not as great as cisplatin. Besides human 2008 ovarian cancer cells we also found that human lung (A549) and head and neck cancer cells (SCC 25) were transiently sensitized by cisplatin. The transfectability of the two commercially available cationic liposomes, Lipofectin and LipofectAmine, was either weak or not consistent among tumors tested. In vivo transfection efficiency of 2008 cells was the highest at 1 microgram DNA per nmol or microgram liposome with all three cationic liposomes. In vitro transfection efficiency of 2008 cells at 1:1 (microgram of DNA:nmole of DC-chol/DOPE liposome) increased in a dose-dependent manner while at 1:10, an optimal ratio for in vitro lipofection, rapidly decreased with an increase in dose. This result indicated that there was a correlation between in vivo and in vitro lipofection at 1:1 ratio for delivering liposomal DNA. Most of the DNA injected into the tumor was concentrated in the tumor and in the skin above the tumor whether cisplatin was preinjected or liposomes were used as carriers.

Topics: Animals; Antineoplastic Agents; Bleomycin; Cation Exchange Resins; Chloramphenicol O-Acetyltransferase; Cholesterol; Cisplatin; DNA; Female; Head and Neck Neoplasms; Lipids; Liposomes; Lung Neoplasms; Mechlorethamine; Mice; Mice, SCID; Ovarian Neoplasms; Paclitaxel; Phosphatidylethanolamines; Transfection